(A) Still image of nascent RB branches expressing EB3-GFP in a live wild type embryo at 23hpf. (A’) Zoom in of the branches outlined in white in (A).(A”) EB3-GFP signal in nascent branches in (A’). (A’’’) Representative kymograph generated from nascent branch outlined in magenta in (A’). (B) Quantification of peripheral axon comet frequency. NS by Mann-Whitney test, p=0.8410. Wild type N=22 nascent branches from 9 embryos, klc4uw314 N=25 nascent branches from 8 embryos. (C-D’’) Representative examples of HNK-1 and acetylated tubulin stained embryos at 24hpf. White arrows in (C’-C’’) indicate nascent branches with tubulin acetylation. Magenta arrows in (D’-D’’) indicate nascent branches lacking tubulin acetylation. (E) Percentage of nascent branches containing acetylated tubulin out of all nascent branches. Fewer nascent branches contained acetylated tubulin in klc4uw314 mutants. ****p<0.0001 by Mann-Whitney test. Wild type N=132 nascent branches from 15 embryos, and klc4uw314 N=143 nascent branches from 16 embryos. Scale bars = 20 μm. Error bars in box and whisker plots represent 5-95th percentile.
